• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

RAB21 Activity Assay Using GST-fused APPL1 Authors:  Steve Jean and Amy A. Kiger, date: 02/20/2016, view: 4355, Q&A: 0
The Rab family of small GTPases are essential regulators of membrane trafficking events. As with other small GTPase families, Rab GTPases cycle between an inactive GDP- bound state and an active GTP-bound state. Guanine nucleotide exchange factors (GEFs) promote Rab activation with the exchange of bound GDP for GTP, while GTPase-activating ...
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Cancer Biology

In vitro Flow Adhesion Assay for Analyzing Shear-resistant Adhesion of Metastatic Cancer Cells to Endothelial Cells Authors:  Shin-Ae Kang, Sandra Bajana and Takemi Tanaka, date: 02/20/2016, view: 7406, Q&A: 1
Hematogenous metastasis is a primary cause of mortality from metastatic cancer. The shear-resistant adhesion of circulating tumor cells to the vascular endothelial cell surface under blood flow is an essential step in cell extravasation and further tissue invasion. This is similar to a process exploited by leukocytes for adhesion to inflamed blood ...
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Cell Biology

VAMP8-3xHA Uptake Assay in HeLa Cells Authors:  Steve Jean and Amy A. Kiger, date: 02/20/2016, view: 4255, Q&A: 0
Transmembrane proteins are rarely exclusively localized to a specific vesicle or an organelle. Most transmembrane proteins undergo complicated trafficking routes. Thus, transmembrane proteins are under constant flux, and at steady state, found on a variety of vesicles or organelles. This characteristic makes the study of their trafficking routes ...
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Immunology

Isolation of Nippostrongylus brasiliensis Larvae from Mouse Lungs Authors:  Mali Camberis, Tiffany Bouchery and Graham Le Gros, date: 02/20/2016, view: 6061, Q&A: 0
The rodent parasite Nippostrongylus brasiliensis (N. brasiliensis) models the salient features of helminth infection including skin penetration, migration from tissues to lung, maturation and egg production in the gut. As a potent activator of systemic and mucosal Th2 immune responses, Nippostrongylus brasiliensis has ...
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Dye Labeling of Live Nippostrongylus brasiliensis Larvae for Visualization in Host Tissue Authors:  Tiffany Bouchery, Mali Camberis and Graham Le Gros, date: 02/20/2016, view: 4623, Q&A: 0
Visualization of the interaction between parasitic nematodes and their host enables a better understanding of the development of the nematode during the infectious stages of its life cycle and of the effects of host response on nematode integrity in tissues. Appropriate live imaging of these nematode/host interactions, to date has been hindered by ...
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Neuroscience

Repeated Cross-fostering Protocol as a Mouse Model of Early Environmental Instability Authors:  Alessandra Luchetti, Marco Battaglia and Francesca R. D’Amato, date: 02/20/2016, view: 4464, Q&A: 0
Early life events have a crucial role in programming the individual phenotype indeed the exposure to traumatic experiences during infancy can increase later risks for a variety of neuropsychiatric conditions, including mood and anxiety disorders. Several studies in rodents demonstrated the impact of short and long sessions of separation/isolation ...
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Synaptoneurosome Preparation from C57BL/6 Striata Authors:  Emma Puighermanal, Anne Biever and Emmanuel Valjent, date: 02/20/2016, view: 6310, Q&A: 0
Activity-dependent local mRNA translation endows synapses to remodel their structure and function (Bramham and Wells, 2007). This process is tightly controlled by the state of phosphorylation of several components of the translational machinery including initiation factors and ribosomal proteins (Buffington et al., 2014). The present ...
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Stem Cell

Immunofluorescent Staining of Mouse Intestinal Stem Cells Authors:  Kevin P. O’Rourke, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 19018, Q&A: 1
Immunofluorescent staining of organoids can be performed to visualize molecular markers of cell behavior. For example, cell proliferation marked by incorporation of nucleotide (EdU), or to observe markers of intestinal differentiation including paneth cells, goblet cells, or enterocytes (see Figure 1). In this protocol we detail a method to fix, ...
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Isolation, Culture, and Maintenance of Mouse Intestinal Stem Cells Authors:  Kevin P. O’Rourke, Sarah Ackerman, Lukas E Dow and Scott W Lowe, date: 02/20/2016, view: 15209, Q&A: 6
In this protocol we describe our modifications to a method to isolate, culture and maintain mouse intestinal stem cells as crypt-villus forming organoids. These cells, isolated either from the small or large intestine, maintain self-renewal and multilineage differentiation potential over time. This provides investigators a tool to culture wild ...
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